LOCUS       sequence-433895-        8183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lysosome-targeted LOVPLD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8183)
  AUTHORS   Li XL, Tei R, Uematsu M, Baskin JM
  TITLE     Ultralow Background Membrane Editors for Spatiotemporal Control of 
            Phosphatidic Acid Metabolism and Signaling.
  JOURNAL   ACS Cent Sci. 2024 Jan 30;10(3):543-554. doi: 
            10.1021/acscentsci.3c01105. eCollection 2024 Mar 27.
  PUBMED    38559292
REFERENCE   2  (bases 1 to 8183)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acscentsci.3c01105"; journalName: "ACS Cent Sci"; date: 
            "2024-01-30- 30"; volume: "10"; issue: "3"; pages: "543-554"
COMMENT     Sequence Label: pCDNA3-p18-mCherry-LOVPLD
FEATURES             Location/Qualifiers
     source          1..8183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1063..1767
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1205..1223)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1473..1492
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3736..3754)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3737..3754)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3774..3791)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3780..4004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4050..4478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4137..4156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4347..4368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4487..4507)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4492..4821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4672..4807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4734..4753
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4888..5682
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4942..4961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5552..5571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5856..5977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5893..5912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5947..5966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6026..6042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6026..6042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6039..6061)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6050..6066
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6074..6080,6081..6098,6099..6104))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6119..6140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6257..6274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6428..7016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6508..6527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7187..7978,7979..8047))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7810..7829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8048..8152)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atgatggggt gctgctacag cagcgagaac gaggactcgg accaggaccg agaggagcgg
      961 aagctgctgc tggaccctag cagcccccct accaaagctc tcaatggagc cgagcccaac
     1021 tacaccggtt ccagtggagg cggtagctca ggttcgggta ccgtgagcaa gggcgaggag
     1081 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1141 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1201 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1261 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     1321 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1381 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1441 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     1501 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     1561 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     1621 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     1681 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     1741 accggcggca tggacgagct gtacaaggga tccgcagatt cagctacacc acatttggac
     1801 gccgttgagc aaacattgag gcaagtgtcc ccaggcctgg agggcgatgt atgggaaaga
     1861 actagtggaa ataaattgga cggaagtgcc gctgatccca gtgattggtt gttgcaaacc
     1921 ccaggatgtt ggggcgatga tagatgcgtt gacagagttg ggacaaaaag gttgctggct
     1981 aagatgactg aaaatatagg gaatgccacc agaaccgtag atattagtac acttgcacct
     2041 ttcccaaacg gcgcattcca agacgctatt gtcgctggtt tgaaggagtc cgcagctaga
     2101 ggcaataagt tgaaagttag gatactggtt ggggctgcac ccgtctacca catgaatgta
     2161 ataccttcca agtataggga cgaattgact gcaaaacttg gcaaagccgc agaaaacata
     2221 actttgaacg tggctagtat gactacatca aaaacagctt tctcctggaa ccatagcaaa
     2281 atcttggttg tagacgggca aagcgccctt actggtggaa taaatagctg gaaggatgat
     2341 tacttggata ctactcatcc agtgtctgat gttgatcttg cactgaccgg accagctgca
     2401 ggctctgccg gtagatacct tgacactttg tggacatgga cctgccaaaa caagagtaac
     2461 atcgcttccg tctggtttgc agcttccggc aacgcagggt gcatggccac tatgcataaa
     2521 gatacaaacc ccaaggcatc acctgccacc gggaatgtcc caataatagc cgttggaggg
     2581 ttgggcgttg gcatcaaaga tgtggacccc aagtccactt tcaggcccga tctgccaaca
     2641 gctagcgata ctaagtgtgt ggtaggactt cacgacaata ctaacgccga attcgagttt
     2701 cgggcaacca cactggaacg gatcgagaaa agcttcgtga ttactgatcc gagactgcct
     2761 gacaacccaa tcatttttgt gagcgattcc ttcctgcagc tgacagaata ttctcgggaa
     2821 gagatcctgg ggcgcaattg ccgttttctg cagggacccg agacagaccg tgccactgtt
     2881 cggaaaatca gagatgctat tgacaaccag actgaagtga ccgttcagct gatcaattat
     2941 accaagagcg gcaagaagtt ctggaacgtg ttccacctgc agccgatgcg cgattataag
     3001 ggcgacgtcc agtacttcat tggcgtgcag ctggatggca ccgaacgtct tcatggcgcc
     3061 gctgagcgtg aggcggtctg cctggtcaaa aagacagcct ttcagattgc tgcggccgct
     3121 gatagagact atgatacagt caacccagaa gagagtgcct tgagagctct tgttgcttca
     3181 gcaaaaagtc acattgaaat atctcagcag gatttgaacg ctacatgtcc acccttgcct
     3241 aggtatgaca ttagattgta cgatgccttg gctgctaaaa tggctgctgg ggttaaggtg
     3301 agaattgtgg ttagcgaccc agccaacagg ggcgcagtgg gcagtgtggg ttacagccaa
     3361 ataaaatcac ttagtgaaat atcagacact ttgagaaaca ggcttgcaaa tatcacaggc
     3421 agtcaacaag ctgccaaaac tgcaatgtgt agcaatttgc agcttgcaac ctttagatcc
     3481 tcccccaacg acaaatgggc ggacgggcat ccttatgccc aacaccataa gttggtttcc
     3541 gtcgactcat ccacatttta tatcggttca aaaaatttgt acccatcctg gctgcaagat
     3601 tttggctaca tagtggaatc cccagaggct gccaagcaac tggatgccaa attgcttgac
     3661 cctcagtgga agtactcaca ggagactgca acagtggatt acgcaagagg aatctgcaac
     3721 gcttaagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     3781 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3841 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3901 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3961 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4021 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4081 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4141 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4201 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4261 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4321 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4381 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4441 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4501 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4561 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4621 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4681 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4741 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4801 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4861 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4921 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4981 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5041 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5101 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5161 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5221 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5281 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5341 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5401 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5461 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5521 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5581 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5641 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5701 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5761 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5821 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5881 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5941 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6001 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6061 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6121 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6181 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6241 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6301 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6361 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6601 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6841 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6901 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7201 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7261 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7321 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7381 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7441 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7501 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7561 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7621 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7681 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7741 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7801 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7861 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7921 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7981 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8041 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8101 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8161 cccgaaaagt gccacctgac gtc
//
